For numerous types of bioanalysis, the sensitive quantitation of a biomolecule at low levels in a sample is highly desirable. For example, it may be desirable to monitor the dynamic expression levels of an intact, post-translationally modified protein in a particular cell or tissue sample or samples. In many cases, the amount of sample of interest; for example, the number of cells or mass of tissue, may be very small. Additionally, the number of copies of the target protein of interest may be very low. In such cases, it may be desirable to assay a protein concentration in sub-femtomole concentrations.
Currently, proximity binding assays as a class of analyses offer the advantages of the sensitivity and specificity of biorecognition binding, along with the exponential signal amplification offered by a variety of oligonucleotide amplification reactions, such as the polymerase chain reaction (PCR).
However, the combination of a binding event, followed by an oligonucleotide amplification reaction event produces data with characteristics requiring specialized analysis methods. Such methods should be readily adapted to the broad class of proximity binding assays, and should provide the user with results presented in readily useful form and format. Accordingly, there is a need in the art for methods for the analysis of proximity binding assay (PBA) data.